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Rice fat synthesis related gene and use thereof

A fat content, amino acid technology, applied in the field of gene technology and botany, can solve the problems of no gene cloning and functional research, no gene production application, etc.

Inactive Publication Date: 2009-03-04
SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, although multiple QTls loci controlling fat content have been identified in recent years, there are no reports of the corresponding genes being successfully cloned and functionally studied, let alone the application of these genes in production

Method used

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  • Rice fat synthesis related gene and use thereof
  • Rice fat synthesis related gene and use thereof
  • Rice fat synthesis related gene and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0202] Example 1 Obtaining of OsLFL1 Mutant Plants

[0203] T-DNA tag isolation of genes is still one of the most important means for the study of plant functional genes. T-DNA (transfer DNA) is a piece of DNA located on the Agrobacterium tumefaciens Ti (tumor inducing) plasmid or Agrobacterium rhizogenes Ri (root inducing) plasmid, which can be transferred from Agrobacterium And it is stably integrated into the plant chromosome, so it can be used as a kind of exogenous gene carrier to transfer exogenous DNA to plant cells and regenerate transgenic plants capable of expressing exogenous genes; As the T-DNA sequence is known, once the genetic analysis confirms that the mutant phenotype is linked to the inserted tag, the flanking sequences at both ends of the tag can be obtained by IPCR or TAIL-PCR, and then The flanking sequences are compared with the genomic sequence to obtain information about the mutant gene. Since Agrobacterium is very effective in transforming heterologo...

Embodiment 2

[0205] Example 2 Cloning of OsLFL1 gene

[0206] Separation of adjacent sequence of T-DNA insertion site by Tail-PCR showed that T-DNA was inserted on rice chromosome 1 and located in the promoter region of OsLFL1 gene locus, 186bp away from the start codon ATG. See figure 1 .

[0207] The rice chromosomal site where the T-DNA was inserted was analyzed by this adjacent sequence. The full-length sequence of the gene was obtained from the rice BAC clone B1142C05 (from the International Rice Genome Sequencing Project Research Organization) according to the comparison analysis, which is SEQ ID NO:1. Using the total RNA extracted from rice, the mRNA fragment of the OsLFL1 gene was isolated and sequenced by 5' and 3' RACE (rapid amplification of ends), and the coding sequence of the OsLFL1 gene, SEQ ID NO: 2, was integrated. The sequence of the OsLFL1 protein is deduced according to the rice universal codon coding rule and the coding sequence SEQ ID NO: 2 of the OsLFL1 gene.

Embodiment 3

[0208] Example 3 Sequence Analysis of OsLFL1 Gene

[0209]The sequence of the OsLFL1 gene was analyzed using the analysis tools "Align two sequences (bl2seq)" and "Protein-protein BLAST (blastp)" in http: / / www.ncbi.nlm.nih.gov / BLAST / . The analysis results are as follows:

[0210] SEQ ID NO: 1 shows that in the OsLFL1 genome sequence, the 1-3915 region is the promoter, the 3916-3918 position is the start codon ATG, the 8734-8736 position is the stop codon TAG, 3268-4428, 5885-5926, 6040 -6126, 6735-6838, 8200-8282, 8362-8978 are exons, 4429-5884, 5927-6039, 6127-6734, 6839-8199, 8283-8361 are introns.

[0211] SEQ ID NO: 2 shows that the 48th-50th position in the coding sequence of the OsLFL1 gene mRNA is the initiation codon ATG, and the 1254-1256th position is the termination codon TAG.

[0212] In SEQ ID NO: 3, positions 177-296 of the OsLFL1 protein coding sequence are B3 DNA binding domains, and positions 296-402 are transcriptional activation domains.

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Abstract

The invention belongs to the field of biotechnology and botany, and discloses a new crop fat synthesis related gene and application thereof. The invention also discloses a method for improving fat content in crop by changing gene expression level or activity. The crop fat synthesis related gene is employed to improve appearance and taste quality of paddy rice, and is introduced into crop such as oil crop, potato, and maize so as to change the fat content of tuber or seed, such as improve fat (oil) content and oil yield in rape seed.

Description

technical field [0001] The invention belongs to the fields of gene technology and botany. Specifically, the present invention relates to a new rice fat synthesis-related gene OsLFL1 and its application. Background technique [0002] Rice (Oryza sativa) is an important cereal crop and the most important food crop in agricultural production in my country. Rice is the main ration of Chinese people. In the past few decades, genetic breeders and agricultural experts in my country have made great contributions to the increase of rice production. In the past, in the breeding work in our country, because the yield was often the first choice, a considerable number of rice varieties had a high yield level, but their internal qualities such as starch, protein, fat and other nutrients were often unbalanced, and their eating quality was such as food quality. Poor mouthfeel and poor oiliness, etc. Because along with the raising of people's living standard, people have proposed new hig...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12N15/29C12N15/82
CPCC07K14/415C12N15/8247
Inventor 张景六彭凌涛李琳沈革志
Owner SHANGHAI INST OF BIOLOGICAL SCI CHINESE ACAD OF SCI
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